STAINING

The process of adding a dye to a

bacterial culture
Dyes
 Basic dye—possess a positive charge
 Acidic dye—possess a negative charge

Remember, bacteria posses a slight

negative charge on their surface
SIMPLE STAINING
 Use only one dye
 For the purpose of viewing bacterial shapes
and arrangements

 Simple Stain the following:

1) E. coli
2) S. aureus
3) A colony from the “Microbes in the
Environment” plate
3 Types of Staining
Procedures
 Simple Staining (shapes and
arrangements)
 Differential Staining (Gram reactions)
 Special Staining (Capsule, flagella,
spores)
PROCEDURE:
 Prepare smear of bacterium
 Air dry
 Heat fix the bacteria to the slide (release
of “sticky” proteins from the cell surface
of the bacteria adheres the bacterial cell
to the slide)
 Apply crystal violet to the smear; let stand
45 seconds
Procedure Cont.
 Rinse with distilled water or tap water
 Blot dry with bibulous paper
 View using microscope
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Page 54
 Methylene blue
Crystal Violet
Fuchsin

Simple stains allow visualization of

Shapes
Arrangements

Proteus vulgaris
Staphyloccocus aureus
Bacillus cereus

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Fracisella tularensis
Causitive agent of Rabbit fever
Methylene blue stain

Sacharromyces cerevisiae
(Brewer’s yeast)
Methylene blue stain
 Bacillus anthracis (anthrax)
Gram positive
Crystal violet stain

Campylobacter jejuni (traveler’s diarrhea)

Gram negative
Fuchsin stain